Obg-Like ATPase 1 Enhances Chemoresistance of Breast Cancer via Activation of TGF-β/Smad Axis Cascades
Obg-Like ATPase 1 Enhances Chemoresistance of Breast Cancer via Activation of TGF-β/Smad Axis Cascades
复制标题
Obg 样 ATP 酶 1 通过激活 TGF-β/Smad 轴级联增强乳腺癌的化疗耐药性
DOI：
10.3389/fphar.2020.00666
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发表时间：
2020-05-27
影响因子：
5.6
通讯作者：
Xiao, Gary Guishan
中科院分区：
文献类型：
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作者：
Liu, Jianzhou;Miao, Xiaoyu;Xiao, Gary Guishan
了解耐药的分子机制有助于确定乳腺癌治疗的有效靶点。在这项研究中，我们研究了Obg样ATP酶1的调节作用，它参与了乳腺癌耐药的多种用途。通过连续增加紫杉醇浓度建立了耐紫杉醇细胞系（MCF-7-PTR）。MTT法用于验证获得性耐药或OLA 1修饰的细胞系。进行qRT-PCR、蛋白质印迹、细胞凋亡和细胞周期测定以评价细胞系中的基因和蛋白质表达。通过一系列的体外实验，研究了OLA 1在乳腺癌中的调控作用。我们证明了OLA 1与乳腺癌的获得性或内在耐药性高度相关。进一步的研究表明，OLA 1的表达增强通过TGF-β/Smad信号通路促进肿瘤细胞EMT过程，导致抗凋亡相关蛋白（切割型caspase 3、Bax、Bcl-2）表达增强，肿瘤细胞微管解聚增强。我们的研究结果表明，OLA 1增强抗凋亡能力，阐明了OLA 1在促进乳腺癌化疗耐药中的调节作用。因此，通过敲低OLA 1可以显著增强疾病的化学敏感性，这导致TGF-β/Smad信号级联的失活，微管聚合，并促进细胞凋亡。我们的数据表明，OLA 1可能被开发为一个潜在的目标，以改善乳腺癌患者的化疗。
Understanding the molecular mechanism of drug resistance helps to identify an effective target for breast cancer therapy. In this study we investigated the regulatory role of Obg-like ATPase 1 which is involved in multiple uses of drug resistance against breast cancer. Paclitaxel resistant cell line (MCF-7-PTR) was developed by a continuous increasing paclitaxel concentration. MTT assay was used to validate either acquired resistant or OLA1 modified cell lines. qRT-PCR, western blotting, apoptosis, and cell cycle assays were executed to evaluate gene and protein expression in cell lines. A series of in vitro assays was performed in the cells with RNAi-mediated knockdown to expound the regulatory function of OLA1 in breast cancer. We demonstrated that OLA1 was highly correlated with either acquired or intrinsic resistance of breast cancer. Further study showed that escalated expression of OLA1 promoted the EMT process in tumor cells through TGF-beta/Smad signaling cascades, resulting in the enhanced expression of anti-apoptosis-related proteins (cleaved caspase3, Bax, Bcl-2) and the strengthening depolymerization of microtubules in tumor cells. Our findings revealed that OLA1 enhanced the anti-apoptotic ability and elucidated a regulatory role of OLA1 in promoting chemotherapy resistance of breast cancer. Chemo-sensitivity of the disease can be thus enhanced significantly by knocked down OLA1, which led to the inactivation of the TGF-beta/Smad signaling cascades, polymerized microtubules, and promoted cell apoptosis. Our data suggest that OLA1 may be developed as a potential target to improve chemotherapy of patients with breast cancer.